Butyric acid matrix metalloproteinase inhibitors

ABSTRACT

Compounds of the formula ##STR1## wherein R 1  and R 2  include hydrogen, alkyl, substituted alkyl, halo, and arylalkyl; R 3  is hydroxy, alkoxy, or hydroxyamino; X is O, S, or NOH; and R 4  and R 5  include hydrogen, alkyl, and aryl are useful for inhibiting matrix metalloproteinase enzymes in animals, and as such, prevent and treat diseases resulting from the breakdown of connective tissues.

This application claims benefit of Provisional Application 60/024,025filed Aug. 16, 1996.

FIELD OF INVENTION

This invention relates to a group of butyric acid derivatives whichinhibit matrix metalloproteinase enzymes and thus are useful fortreating diseases resulting from tissue breakdown, such as arthritis andosteoporosis.

BACKGROUND OF THE INVENTION

Matrix metalloproteinases are naturally occurring enzymes found in mostmammals and are associated with the breakdown of connective tissues. Theclass includes gelatinase A and B, stromelysin-1, fibroblastcollagenase, neutrophil collagenase, matrilysin, and other forms ofcollagenase. These enzymes have been implicated with a number ofdiseases which result from breakdown of connective tissue, such asrheumatoid arthritis, osteoarthritis, osteoporosis, periodontitis,multiple sclerosis, gingivitis, corneal epidermal and gastriculceration, atherosclerosis, neointimal proliferation which leads torestenosis and ischemic heart failure, and tumor metastasis. A methodfor preventing and treating these diseases is now recognized to be byinhibiting metalloproteinase enzymes, thereby curtailing and eliminatingthe breakdown of connective tissues that results in the disease states.

Several inhibitors of metalloproteinases have been identified. Manyinhibitors are complex peptides, for instance as described by Chapman,et al., in J. Med. Chem., 1993;36:4293-4301. Small peptide inhibitorsare also known, for example as described in U.S. Pat. Nos. 4,599,361 and5,270,326, as well as nonpeptides as in WO 95/35276.

The need continues for small molecular weight molecules which can beeconomically prepared and yet are effective inhibitors ofmetalloproteinases. We have now discovered a group of butyric acidderivatives which have exceptionally good inhibitory activity. An objectof this invention is to provide such compounds, their pharmaceuticalformulations, and a method for using them to treat diseases mediated bymetalloproteinases.

SUMMARY OF THE INVENTION

This invention provides butyric acid derivatives which are inhibitors ofmatrix metalloproteinases. The invention compounds have Formula I##STR2## wherein:

X is O, NOR₉, S, OH, SH, or ##STR3##

R₇ and R_(7a) independently are

hydrogen,

C₁ -C₂₀ alkyl or substituted C₁ -C₂₀ alkyl,

(CH₂)₀₋₆ -aryl,

(CH₂)₀₋₆ -heteroaryl, or

(CH₂)₀₋₆ -cycloalkyl;

R₁ and R₂ independently are

hydrogen,

C₁ -C₂₀ alkyl or substituted C₁ -C₂₀ alkyl,

halo,

NO₂,

CN,

CHO,

COR₆,

COOR₆,

SO₃ R₆,

OR₆,

CONR₄ R₅,

(CH₂)₀₋₆ -aryl,

(CH₂)₀₋₆ -heteroaryl, or

(CH₂)₀₋₆ -cycloalkyl;

R₆ is hydrogen,

C₁ -C₂₀ alkyl or substituted C₁ -C₂₀ alkyl;

aryl is phenyl or substituted phenyl;

R₃ is hydroxy,

O--C₁ -C₂₀ alkyl or substituted O--C₁ -C₂₀ alkyl,

O--(CH₂)₁₋₃ aryl, or

NHOR₆ ;

R₄ and R₅ independently are hydrogen,

C₁ -C₂₀ alkyl or substituted C₁ -C₂₀ alkyl,

C₂ -C₂₀ alkenyl or substituted C₂ -C₂₀ alkenyl,

(CH₂)₀₋₆ -aryl,

(CH₂)₀₋₆ -(O or S)-aryl,

(CH₂)₀₋₆ -heteroaryl,

(CH₂)₀₋₆ -(O or S)-heteroaryl;

or one of R₄ and R₅ is hydrogen and the other is:

COR₈,

CSR₈,

CONR₈ R₉,

CSNR₈ R₉,

COOR₈,

COSR₈, ##STR4## S(O)₁ or 2 -C₁ -C₂₀ alkyl or substituted alkyl, S(O)₁ or2 -aryl

S(O)₁ or 2 -heteroaryl, or

S(O)₁ or 2 -cycloalkyl;

Y is ##STR5##

R₈ and R₉ independently are

hydrogen,

C₁ -C₂₀ alkyl or substituted C₁ -C₂₀ alkyl,

C₂ -C₂₀ alkenyl or substituted C₂ -C₂₀ alkenyl,

(CH₂)₀₋₆ -(O or S)₀₋₁ -aryl,

(CH₂)₀₋₆ -(O or S)₀₋₁ -heteroaryl, or

(CH₂)₀₋₆ -(O or S)₀₋₁ -cycloalkyl;

W, W¹, Z, and Z¹ independently are CR₁ or N;

the term ##STR6## is a double bond when X is O, S, NOR₉, or

N-NR₇ R_(7a), and a single bond when X is OH or SH; and thepharmaceutically acceptable salts, isomers, stereoisomers, and solvatesthereof.

Preferred compounds are defined by Formula II ##STR7## wherein R₁, R₂,R₃ R₄, R₅, and X are as defined above.

Further preferred compounds have the formula ##STR8## and especiallypreferred are those wherein X is O, and R₃ is hydroxy. The mostpreferred compounds of the invention are in this latter group where R₄is hydrogen, and R₅ is COR₈, COOR₈, or CONR₈ R₉, where R₈ and R₉ arealkyl or substituted alkyl, aryl or substituted aryl.

A further embodiment of this invention is a pharmaceutical formulationcomprising a compound of Formula I admixed with a pharmaceuticallyacceptable carrier, excipient, or diluent therefor.

The invention additionally provides a method for inhibiting thehydrolytic activity of a matrix metalloproteinase enzyme comprisingadministering to a mammal an effective amount of a compound of FormulaI. In a preferred embodiment, the invention is a method for inhibitingor treating multiple sclerosis, atherosclerotic plaque rupture, aorticaneurysms, heart failure, restenosis, periodontal disease, cornealulceration, cancer metastasis, tumor angiogenesis, arthritis, and forwound healing.

DETAILED DESCRIPTION OF THE INVENTION

As used herein, the terms "C₁ -C₂₀ alkyl" and "substituted C₁ -C₂₀alkyl" mean straight or branched aliphatic groups having from 1 to 20carbon atoms, preferably 1 to 12, and typically 1 to 6, optionallysubstituted. Exemplary alkyl groups include methyl, ethyl, isopropyl,1,1-dimethylheptyl, n-decyl, 1-n-butyl-2-isopropyldecyl, tert-butyl,isoundecyl, and the like. Substituted alkyl groups are the foregoing C₁-C₂₀ alkyl groups having one or more, typically from 1 to 3, substituentgroups such as halo, hydroxy, C₁ -C₄ alkoxy, thio, C₁ -C₄ alkylthio,phenyl, alkylphenyl, phenoxy, alkylphenoxy, substituted phenyl orsubstituted phenoxy. Typical "substituted C₁ -C₂₀ alkyl" groups thusinclude hydroxymethyl, n-butoxymethyl, chloromethyl,1-(3-chlorophenoxy)ethyl-, 1,2,4-trifluorohexyl, 1-(phenoxymethyl)ethyl,2-methoxydecyl, 4-methylthio-5-bromo-undecyl, 1-ethyl-3-phenylbutyl,4-(3,4-dibromophenyl)-heptyl, pentadecafluoro-octyl, and2,2,2-trifluoroethyl.

The terms "C₂ -C₂₀ alkenyl" and "substituted C₂ -C₂₀ alkenyl" mean theforegoing alkyl groups having one to four non-adjacent double bonds.Examples include prop-2-enyl, but-2-enyl, oct-2,4-dienyl,2-phenylethenyl, 3-naphthyl-but-2-enyl, and 6-(2-pyridyl)-hex-3-enyl.

The term "halo" includes fluoro, bromo, chloro, and iodo. Several of theR groups in Formula I can be a group defined as "COR₆ ", "COOR₆ ", "SO₃R₆ ", and "OR₆ ", where R₆ is hydrogen, alkyl, or substituted alkyl asthose terms are defined above. The COR₆ groups thus include formyl,acetyl, propionyl, pivaloyl, 3-hydroxybutyryl, undecanoyl,4-n-butyl-5-chloro-6-methylthio-heptanoyl, 4-bromophenoxyacetyl, and4-phenylpentanoyl. Typical COOR₆ groups include methoxycarbonyl,benzyloxycarbonyl, n-octyloxycarbonyl,5-methoxy-6-thioheptyloxycarbonyl, and the like. Typical SO₃ R₆ groupsinclude methoxysulfonyl, 3-chlorobenzyloxysulfonyl, and4-chlorobutyloxy-sulfonyl. The OR₆ groups include methoxy, tert-butoxy,benzyloxy, 4-nitrobenzyloxy, and the like.

The term "CONR₄ R₅ " means an aminocarbonyl substituent such asmethylaminocarbonyl, benzylaminocarbonyl, diethylaminocarbonyl,3-pyridylmethylaminocarbonyl, and the like.

The term "aryl" means a monocyclic or bicyclic aromatic hydrocarbylgroup, for example, phenyl or naphthyl, optionally substituted with fromone to five groups, typically one, two, or three, selected from halo,hydroxy, methylenedioxy, phenyl, nitro, cyano, C₁ -C₆ alkyl, C₁ -C₆alkoxy, thio, C₁ -C₆ alkylthio, amino, C₁ -C₆ alkyl and dialkylamino,CHO, COR₆, COOR₆, OR₆, SO₃ R₆, or CONR₄ R₅. Typical (CH₂)₀₋₆ -arylgroups thus include phenyl, 3-hydroxyphenyl, 2-methylphenyl,4-chlorobenzyl, 3,4-methylenedioxyphenyl, 4-(2-methylthiophenyl)butyl,2-(3-aminophenyl)ethyl, 6-(2-chloro-3-methylamino-4-formylphenyl)hexyl,naphthyl, 3-chloronaphthyl, and 2,4,6-trimethyl-naphthyl.

The term "(CH₂)₀₋₆ -(O or S)-aryl" means the foregoing aryl groupsbonded through oxygen or sulfur. Examples include phenoxymethyl,2-phenylthioethyl, l-naphthyloxymethyl, and3-bromo-l-naphthylthiomethyl.

The term "(CH₂)₀₋₆ -heteroaryl" means a heteroaryl group bonded directlyor through from one to six methylene groups. The term "heteroaryl" meansa monocyclic or fused bicyclic aromatic ring having from 4 to 12 carbonatoms and from 1 to 3 heteroatoms selected from O, S, or N. Theheteroaryl group can be substituted with the same groups as indicatedabove for phenyl. Typical heteroaryl groups thus include pyridyl, 2- or3-furanyl, 2- or 3-thienyl, 2- or 3-pyranyl, 2-, 4-, or 5-pyrimidinyl,3- or 4-pyridazinyl, 2- or 3-morpholinyl, indolyl, 3-phenyl-2-pyridyl,benzoxazolyl, benzopyranyl, quinolinyl, quinoxalinyl, isoquinolinyl, andpyridopyrimidinyl. Substituted heteroaryl groups include3-chlorothiophene, 3-hydroxyfuryl, and 3-nitromorpholine.

The term "(CH₂)₀₋₆ -(O or S)-heteroaryl" means the foregoing heteroarylgroups bonded through oxygen or sulfur- Examples include3-pyridyloxymethyl, 2-(2-pyranyl)oxyethyl, and 3-furanylthiomethyl.

The term "(CH₂)₀₋₆ -(O or S)₀₋₁ -cycloalkyl" means a cycloalkyl groupbonded directly or through from one to six methylene groups, andoptionally through oxygen or sulfur. "Cycloalkyl" means a monocyclic orbicyclic carbocyclic group having from three to ten carbon atoms, andoptionally substituted with one, two, or three groups selected fromhalo, hydroxy, phenyl, nitro, cyano, C₁ -C₆ alkyl, C₁ -C₆ alkoxy, thio,C₁ -C₆ alkylthio, amino, C₁ -C₆ alkyl and dialkylamino, CHO, COR₆,COOR₆, OR₆, SO₃ R₆, and CONR₄ R₅. Typical cycloalkyl groups thus includecyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, 2-ethylcyclopentyl,3,4-dichlorocyclohexyl, decalinyl, adamantyl, 3-aminocyclooctyl, and2-phenylcyclohexyl.

R₄ and R₅ are nitrogen substituents and include hydrogen, alkyl andsubstituted alkyl, (CH₂)₀₋₆ -(O or S)₀₋₁ -aryl and (CH₂)₀₋₆ -(O or S)₀₋₁-heteroaryl, as those terms are defined above. Preferred compounds ofthe invention are those wherein R₄ is hydrogen, and R₅ is other thanhydrogen, especially COR₈, CSR₈, CONR₈ R₉, CSNR₈ R₉, COOR₈, SO₂ -alkyl,SO₂ -aryl, and COSR₈. These groups define compounds which are,respectively, amides, thioamides, ureas, thioureas, carbamates,sulfonamides, and thiocarbamates. Typical "COR₈ " groups thus includeformyl, acetyl, bromoacetyl, trifluoroacetyl, cyclobutylformyl,3-nitropropionyl, 4-methoxycarbonylbutyryl, benzoyl, 4-phenylbenzoyl,phenoxyacetyl, l-phenoxyethyl, 6-dimethylaminocarbonyl-octanoyl, and4-phenyldecanoyl. Typical "CSR₈ " groups include methylthiocarbonyl,ethylthiocarbonyl, and tert-butylthiocarbonyl. Examples of "CONR₈ R₉ "groups include aminocarbonyl, dimethylaminocarbonyl,dibenzylaminocarbonyl, and the like. The invention includes thioureas,i.e., compounds of Formula I where R₄ is hydrogen, and R₅ is ##STR9##Typical of such R₅ groups are aminothiocarbonyl,diethylaminothiocarbonyl, and benzylaminothiocarbonyl. The inventionalso includes sulfonamides, for example when R₅ is --SO₂ alkyl or --SO2aryl. Typical groups include isopropylsulfonyl, phenylsulfonyl,naphthylsulfonyl, and 4-chlorophenylsulfonyl.

R₄ and R₅ can include "COOR₈ " and "COSR₈ "; preferably, one of R₄ or R₅is hydrogen. Such groups form carbamates and thiocarbamates,respectively. Typical COOR₈ groups include methoxycarbonyl,3-hydroxybutoxycarbonyl, benzyloxycarbonyl,2,4-dibromobenzyloxycarbonyl,l-methoxy-2-phenyl-3-chloropropoxycarbonyl, and10-methoxyundecyloxycarbonyl. Examples of "COSR₈ " groups includemethylthiocarbonyl, benzylthiocarbonyl, and the like.

Ideally, one of R₄ and R₅ is hydrogen when the other is ##STR10## Thesegroups are residues of naturally occurring as well as unnatural aminoacids, for example, glycyl, alanyl, N-methylvalyl, N-benzylleucyl,N,N-dimethylphenylalanyl, threonyl, arginyl, glutamyl, lysyl, tyrosyl,2-amino-6-chlorohexanoyl, 2-benzylamino-3,4-dimethoxyheptanoyl, and thelike.

Ideally, R₄ is hydrogen when R₅ is a group having the formula ##STR11##Examples of these groups include methylaminoformamidocarbonyl,ethoxyformamidocarbonyl, methylthioformamidocarbonyl,diethylaminosulfonylaminocarbonyl, andN-benzyloxysulfonyl-N-ethylaminocarbonyl.

Typical compounds provided by this invention are illustrated by thefollowing general formulas: ##STR12##

Especially preferred compounds are those illustrated below: ##STR13##where R₁₀ is:

--CF₃, ##STR14##

C₁ -C₁₂ alkyl, or perfluoro-C₁ -C₁₂ alkyl, ##STR15##

The invention compounds of Formula I are prepared by any of severalprocedures, utilizing reaction processes well-known to those havingordinary skill in the art of organic chemistry. A typical inventioncompound can be prepared by acylating a tricyclic reactant with an aminosubstituted butyryl acylating agent such as aspartic acid anhydride, ora butyryl agent of the formula ##STR16## where L is a leaving group suchas chloro or formyloxy, and R₃, R₄, and R₅ are as defined above. Theacylation is conducted under Friedal Crafts conditions, for exampleutilizing aluminum chloride as a catalyst, and conducting the reactionin an organic solvent, preferably a halogenated hydrocarbon such asdichloromethane or chloroform. A preferred process comprises reacting atricyclic starting material with an N-substituted D- or L-aspartic acidanhydride according to the following general scheme: ##STR17## Thereactants are combined in approximately equimolar quantities in asolvent such as dichloromethane with an equivalent or excess of aluminumchloride. The acylation typically is complete within about 8 to about 24hours when carried out at a temperature of about -5° C. to about 20° C.The product is readily isolated by pouring the reaction mixture intoice/water, and separating the organic layer. Removal of the solvent byevaporation affords the invention compound, which can be purified bystandard techniques such as chromatography and crystallization.

The compound thus prepared can be converted to other invention compounds(i.e., where one or both of R₄ and R₅ are other than hydrogen, or wherethe 4-oxo group is converted to an oxime) by conventional means. Forexample, a compound of the above formula wherein R₄ is hydrogen, and R₅is an acyl group such as trifluoroacetyl readily reacts with ammonia inan alcohol such as methanol to afford the corresponding primary amine,where both R₄ and R₅ are hydrogen. The primary amine can be acylated oralkylated by conventional means, for example by reaction with an acylhalide such as acetyl chloride or glycyl bromide, a sulfonyl halide suchas isopropylsulfonyl chloride or benzylsulfonyl bromide, an anhydridesuch as acetic anhydride or acetic-formic anhydride, or an alkyl halidesuch as methyl bromide or ethyl iodide, to provide the correspondingN-acyl or N-alkyl derivative. The butanoic acid esters of the invention,i.e., where R₃ is alkoxy or substituted alkoxy, are readily prepared byreacting the free acid of the above formula with an alcohol under acidicconditions, or by other conventional esterification techniques. Suchreactions are depicted in Scheme 1. ##STR18##

The butyric acid esters of Formula I, compounds wherein R₃ is O-alkyl orO-substituted alkyl, are especially useful as intermediates, in thatother sites in the molecule can be derivatized, and the ester functioncan subsequently be hydrolyzed to provide the free acid. The estergroups are readily hydrolyzed to the corresponding carboxylic acids byroutine methods, for instance by reaction with a strong acid such astrifluoroacetic acid, polyphosphoric acid, sulfuric acid, or the like.The hydrolysis generally is carried out at a temperature of about 0° C.to about 25° C., and normally is complete within about 2 to 24 hours.The product, a compound of Formula I wherein R₃ is OH, can be isolatedby diluting the reaction mixture with water and extracting the productinto a water immiscible solvent such as ethyl acetate, dichloromethane,or the like, and then removing the organic solvent, for example byevaporation under reduced pressure. The free carboxylic acids thusformed can be converted to salts by reaction with a base such as sodiumhydroxide, calcium carbonate, or the like. The carboxylic acids also canbe reacted with hydroxylamine hydrochloride to form the correspondinghydroxamic acids, i.e., compounds of Formula I where R₃ is NHOH.

The invention compounds contain at least one asymmetric carbon atom, andas such exist as optically active isomers. The invention contemplatesthe racemic forms as well as the individual isomers. The individualisomers can be prepared from optically pure starting materials, forexample by utilizing naturally occurring amino acids, or by resolvingthe racemate by normal techniques such as chromatography and the like.

Synthesis of various invention compounds of Formula I can be facilitatedby utilizing common protecting groups on functional groups such ashydroxy, carboxy, and amino. Protecting groups will be utilized toprevent unwanted side reactions and are readily removed when desired toprovide the invention compound. The use of protecting groups iswell-documented, for example, by Greene and Wuts in "Protective Groupsin Organic Synthesis," 2nd Ed., 1991, John Wiley & Sons, Inc. Typicalcarboxy acid and hydroxy protecting groups are readily removable esterand ether forming groups such as 2,2,2-trichloroethyl, benzyl, methyl,trimethylsilyl, acetyl, and the like. Primary amino groups, for example,compounds of Formula I wherein R₄ and R₅ both are hydrogen, are readilyprotected with common acyl groups such as acetyl, pivaloyl,trifluoroacetyl, trimethylsilyl, and t-butyldimethylsilyl.

A key intermediate for the synthesis of most of the compounds of thisinvention is the primary amines of Formula I wherein R₄ and R₅ both arehydrogen. These compounds are prepared as described above and inScheme 1. In a preferred embodiment, the key intermediate has theformula ##STR19## where R₁, R₂, and R₃ are as defined above. Suchprimary amino compounds readily undergo standard reactions of amines toprovide invention compounds of Formula I wherein one or both of R₄ andR₅ are other than hydrogen. For example, the primary amines react withacylating agents such as acid halides and acid anhydrides to produce thecorresponding amides. Typical acid halides that can be utilized include##STR20## and typical anhydrides include ##STR21## Ureas, e.g., FormulaI where R₄ is hydrogen and R₅ is CONR₈ R₉, can be prepared by reactingthe primary amine with an isocyanate of the formula R₈ --N═C═O, followedby alkylation or acylation to afford compounds wherein R₉ is other thanhydrogen. Carbamates and thiocarbamates, e.g., Formula I where R₄ ishydrogen and R₅ is ##STR22## respectively, are prepared by reacting theprimary amine with acid halides of the formula ##STR23## Similaracylations can be carried out with acid halides such as ##STR24## Theprimary amines (Formula I where R₄ and R₅ both are H) can also bereacted with chlorocarbonyl isocyanate to give an acid chlorideintermediate that can then be reacted with amines, alcohols, and thiolsto provide invention compounds. The reaction of the amine andchlorocarbonyl isocyanate is depicted as follows: ##STR25## The primaryamine and chlorocarbonyl isocyanate generally are mixed in equimolarquantities in an unreactive organic solvent such as diethyl ether ordichloromethane. The reaction generally is complete within about 1 to 2hours when carried out at about 0° C. The intermediate acid chloridereadily reacts with amines, alcohols, and thiols according to thefollowing scheme: ##STR26## The primary amine (Formula I where R₄ and R₅both are hydrogen) can also be reacted with chlorosulfonyl isocyanate toprovide a sulfonyl chloride: ##STR27## The sulfonyl chloride can bereacted with an amine or alcohol as follows: ##STR28## The primaryamines can additionally be acylated by reaction with a carboxy acid inthe presence of a peptide coupling agent. Typical coupling agentsinclude N,N'-dicyclohexycarbodiimide, N,N'-diisopropylcarbodiimide,N,N'-diethylcarbodiimide, and 1,2-dihydroquinoline.

As noted above, the invention includes pharmaceutically acceptable saltsand solvates of compounds having Formula I. The carboxylic acids (i.e.,R₃ is OH) readily react with inorganic and organic bases to form suchsalts. Typical inorganic bases commonly employed include sodiumhydroxide, potassium carbonate, calcium phosphate, and sodiumbicarbonate. Organic bases routinely used include diethylamine,pyridine, benzylamine, triethanolamine, morpholine, and ethylenediamine.Compounds of Formula I which have a basic group, for example when R₄ andR₅ both are hydrogen, can form acid addition salts by reaction withinorganic or organic acids. Typical acids include hydrochloric acid,sulfuric acid, phosphoric acid, benzoic acid, methanesulfonic acid,p-toluenesulfonic acid and the like. The salts of the inventiontypically are highly crystalline under normal conditions and can bepurified by crystallization from common solvents such as ethyl acetate,chloroform, ethanol, water, toluene, diethyl ether, and hexane. Suchcrystallizations can produce the invention compound as a solvate, forexample as a hydrate or ethanolate, which forms can be utilized inpharmaceutical preparations.

The synthesis of invention compounds is further illustrated in Scheme 2,as well as in the following detailed examples. The examples areillustrative only and are not intended to be limiting in any respect.##STR29##

EXAMPLE 1

(S)-4-Dibenzofuran-2-yl-4-oxo-2-(2,2,2-trifluoroacetylamino)-butyricacid

To a dichloromethane (50 mL) suspension of aluminum chloride (3.19 g,0.024 mol) at 0° C., under an inert nitrogen atmosphere was added via asolid addition funnel an intimate mixture ofN-trifluoroacetyl-L-aspartic acid anhydride (3.01 g, 0.0142 mol) anddibenzofuran (2 g, 0.0119 mol). The resulting brick-red suspension wasallowed to warm to room temperature over 12 hours. The reaction mixturewas poured into ice water (250 mL), and the resulting colorless biphasicmixture was extracted with ethyl acetate (4×100 mL). The combinedorganic extracts were washed with water (2×100 mL), brine (2×100 mL) anddried over magnesium sulfate. The mixture was filtered, and the organicsolution was concentrated by evaporation of the solvent in vacuo toyield a white solid. The solid was recrystallized from 10% ethylacetate/hexane (v/v) to give(S)-4-dibenzofuran-2-yl-4-oxo-2-(2,2,2-trifluoroacetylamino)-butyricacid (2.16 g, 48%) as a while solid; mp 174-176° C.

EXAMPLE 2

(R)-4-Dibenzofuran-2-yl-4-oxo-2-(2,2,2-trifluoroacetylamino)-butyricacid

The general procedure of Example 1 was repeated, utilizingN-(trifluoroacetyl)-D-aspartic acid anhydride to provide(R)-4-dibenzofuran-2-yl-4-oxo-2-(2,2,2-trifluoroacetylamino)-butyricacid; mp 140-144° C.

EXAMPLE 3

(S)-2-Amino-4-dibenzofuran-2-yl-4-oxo-butyric acid

Methanol (40 mL) was saturated with gaseous ammonia at room temperature.To this solution was added(S)-4-dibenzofuran-2-yl-4-oxo-2-(2,2,2-trifluoroacetylamino)-butyricacid (0.5 g, 0.0013 mol) obtained above in Example 1. The reactionmixture was stirred at room temperature for 24 hours and thenconcentrated to dryness in vacuo. The residue was recrystallized fromboiling methanol to yield the title product as a white solid (0.37 g,quant); mp 182-187° C.

EXAMPLE 4

(S)-2-Acetylamino-4-dibenzofuran-2-yl-4-oxo-butyric acid

A suspension of (S)-2-amino-4-dibenzofuran-2-yl-4-oxo-butyric acid(0.025 g, 0.000082 mol) obtained above in Example 3 in acetic anhydride(2 mL) and water (4 mL) was sonicated (Cole Parmer 8850 Ultrasound Bath)for 15 minutes at room temperature. The resulting solution was thenconcentrated in vacuo, and the resulting residue was recrystallized fromethyl acetate/diethyl ether (50:50, v/v) to yield the title product as awhite solid; mp 166-170° C.

EXAMPLE 5

(S)-4-Dibenzofuran-2-yl-2-[3-(2,6-diisopropyl-phenyl)-ureido]-4-oxo-butyricacid

Step (a)(S)-4-Dibenzofuran-2-yl-4-oxo-2-(2,2,2-trifluoro-acetylamino)-butyricacid tert-butyl ester

To a DMF solution (10 mL) of(S)-4-dibenzofuran-2-yl-4-oxo-2-(2,2,2-trifluoroacetylamino)-butyricacid (0.3 g, 0.00079 mol) at 40° C. was added carbonyldimidazole (0.13g, 0.00079 mol). This mixture was stirred for 1 hour and then1,8-diazabicyclo-[5.4.0]undec-7-ene (DBU, 0.12 mL, 0.00079 mol) andt-butanol (0.15 mL, 0.00158 mol) were added. The reaction mixture wasstirred for 24 hours at 40° C., cooled to 24° C., and then diluted withether (20 mL). The solution was washed with water (2×10 mL), brine (2×10mL), and dried over anhydrous magnesium sulfate. The mixture wasfiltered and the filtrate was concentrated to an oil by evaporation ofthe solvent in vacuo. The oil was flash chromatographed on silica geleluting with 20% ethyl acetate/hexane (v/v) to yield the title compound;mp 92-94° C.

Step (b)(S)-4-Dibenzofuran-2-yl-2-[3-(2,6-diisopropoyl-phenyl)-ureido]-4-oxo-butyricacid tert-butyl ester

Into a methanolic solution (100 mL) of the compound from Step (a) above(0.22 g, 0.000505 mol) was bubbled ammonia gas at room temperature for 5minutes. The reaction mixture was stirred at room temperature for 18hours. The solution was then concentrated and flash chromatographed onsilica gel eluting with ethyl acetate to give(S)-2-amino-4-dibenzofuran-2-yl-4-oxo-butyric acid, t-butyl ester (0.112g, 65%) which was used in the next step without further purification.

To a stirred ethyl acetate solution (5 mL) of the product from Step (b)(0.112 g, 0.00033 mol) was added 2,6-diisopropylphenylisocyanate (0.067g, 0.00033 mol) at room temperature under an inert nitrogen atmosphere.The reaction mixture was stirred for 48 hours after which time thesolution was concentrated in vacuo and flash chromatographed on silicagel eluting with 50% ethyl acetate/hexane (v/v) to yield(S)-4-dibenzofuran-2-yl-2-[2,6-diisopropylphenyl)-ureido]-4-oxo-butyricacid tert-butyl ester (0.05 g, 28%); mp 94-97° C.

Step (c)(S)-4-Dibenzofuran-2-yl-2-[3-(2,6-diisopropyl-phenyl)-ureido]-4-oxo-butyricacid

To a dichloromethane solution (2 mL) of the product obtained above inStep (b) (0.05 g, 0.000092 mol) was added trifluoroacetic acid (1 mL)and one drop of anisole at room temperature with stirring. After 7hours, the solution was concentrated and the residue triturated withdiethyl ether. The solid obtained was filtered and air dried to yield(S)-4-dibenzofuran-2-yl-2-[3-(2,6-diisopropyl-phenyl)-ureido]-4-oxo-butyricacid (0.025 g, 56%); mp 205-210° C. (dec).

EXAMPLE 6

(S)-2-Benzoylamino-4-dibenzofuran-2-yl-4-oxo-butyric acid

Step (a) (S)-2-Amino-4-dibenzofuran-2-yl-4-oxo-butyric acid, methylester, hydrochloride

Into a methanolic solution (250 mL) of(S)-4-dibenzofuran-2-yl-4-oxo-2-(2,2,2-trifluoroacetylamino)-butyricacid (1.75 g, 0.0046 mol) was bubbled gaseous hydrogen chloride for 3hours at room temperature. The solution was then concentrated in vacuo,and the resulting solid was triturated with a mixture of ethyl acetateand diethyl ether (1:1, v/v) to yield the title compound (1.22 g, 77%);

mp 156-160° C.

Step (b) (S)-2-Benzoylamino-4-dibenzofuran-2-yl-4-oxo-butyric acidmethyl ester

To a THF/water (1:1, 10 mL) solution of the material obtained above inStep (a) (0.5 g, 0.0015 mol) was added triethylamine (0.22 mL, 0.0015mol). The resulting suspension was stirred at room temperature for 30minutes, at which time more triethylamine (0.22 mL, 0.0015 mol) andbenzoyl chloride (0.21 g, 0.0015 mol) were added. The reaction mixturewas stirred at room temperature for 1 hour and then diluted with 50 mLof water. The solution was extracted with ethyl acetate (2×50 mL). Theorganic extract was washed with water (2×50 mL) and brine (2×50 mL) anddried over anhydrous magnesium sulfate. The solution was filtered andthe filtrate was concentrated in vacuo. Flash chromatography on silicagel eluting with 50% ethyl acetate/hexane (v/v) gave(S)-2-benzoylamino-4-dibenzofuran-2-yl-4-oxo-butyric acid methyl ester(0.39 g, 65%); mp 65-70° C.

Step (c) (S)-2-Benzoylamino-4-dibenzofuran-2-yl-4-oxo-butyric acid

To a dioxan/water solution (1:1, 30 mL) of the material obtained abovein Step (b) (0.32 g, 0.000797 mol) was added lithium hydroxidemonohydrate (0.07 g, 0.00168 mol). The resulting suspension was stirredat room temperature for 4 hours. The reaction mixture was concentratedin vacuo and the residue re-dissolved in water (10 mL) and cooled in anice bath. The cold solution was acidified to pH 2.0 with aqueous 1N HCl.The precipitate which formed was collected by filtration and air driedto yield (S)-2-Benzoylamino-4-dibenzofuran-2-yl-4-oxo-butyric acid (0.29g, 94%); mp 124-130° C.

Following the general procedure of Example 6, the following compoundswere obtained:

EXAMPLE 7

(S)-4-Dibenzofuran-2-yl-4-oxo-2-phenylacetylamino-butyric acid;

mp 94-98° C.

EXAMPLE 8

(S)-4-Dibenzofuran-2-yl-4-oxo-2-(3-phenylpropionylamino)-butyric acid;

mp 118-121° C.

EXAMPLE 9

(S)-4-Dibenzofuran-2-yl-4-oxo-2-(7-phenylheptanoylamino)-butyric acid;

mp 105-108° C.

EXAMPLE 10

(S)-2-[(Biphenyl-4-carbonyl)-amino]-4-dibenzofuran-2-yl-4-oxo-butyricacid

¹ H NMR (DMSO-D₆): δ 8.9 (s, 1H), 8.8 (m, 1H), 8.3-7.4 (m, 16H), 5.1 (m,1H), 3.8 (m, 2H) ppm.

EXAMPLE 11

(S)-4-Dibenzofuran-2-yl-4-oxo-2-(octanoylamino)-butyric acid;

mp 57-60° C.

EXAMPLE 12

(S)-4-Dibenzofuran-2-yl-4-oxo-2-(dodecanoylamino)-butyric acid;

mp 85-88° C.

EXAMPLES 13-14

Following the general procedure of Example 6, except substituting asulfonyl chloride for the benzoyl chloride utilized in Step (b), thefollowing sulfonamides were prepared:

(S)-4-Dibenzofuran-2-yl-4-oxo-2-methanesulfonylamino-butyric acid; mp161-168° C.;

(S) -4-Dibenzofuran-2-yl-4-oxo-2-(4-methylphenylsulfonylamino)-butyricacid;

¹ H NMR (DMSO-d₆): δ 8.9 (s, 1H), 7.2-8.3 (m, 11H), 4.5 (m, 1H), 3.6 (m,2H), 2.8 (m, 1H), 2.2 (s, 3H) ppm.

EXAMPLE 15

By following the procedure of Example 1, except using fluorene insteadof dibenzofuran, there was obtained the compound(S)-4-(9H-fluoren-2-yl)-4-oxo-2-(2,2,2-trifluoroacetylamino)-butyricacid;

mp 111-115° C.

EXAMPLE 16

(S)-4-Dibenzofuran-2-yl-4-hydroxyimino-2-(2,2,2-trifluoroacetylamino)-butyricacid

To a solution of(S)-4-dibenzofuran-2-yl-4-oxo-2,2,2-trifluoroacetylamino)-butyric acid(1.0 g, 0.0026 mol) from Example 1 in 50 mL of methanol was added sodiumacetate trihydrate (1.08 g, 0.0079 mol). The solution was stirred at 24°C. while an aqueous solution (5 mL) of hydroxylamine hydrochloride (0.37g, 0.00527 mol) was added in one portion. The reaction mixture wasstirred for 3 hours at 24° C., and then heated to reflux for 2 hours.The solution was cooled to 24° C. and concentrated to dryness byevaporation under reduced pressure. The solid was rinsed with 10 mL ofwater, and crystallized from ethyl acetate and hexane to afford 0.13 gof(S)-4-dibenzofuran-2-yl-4-hydroxyimino-2-(2,2,2-trifluoroacetylamino)-butyricacid; mp 128-130° C.

EXAMPLE 17

(S)-2-Benzoylamino-4-dibenzofuran-2-yl-4-oxo-butyric acid methyl ester,prepared as described in Step (b) of Example 6, was dissolved intetrahydrofuran containing one equivalent of triethylamine. The solutionwas stirred at room temperature for 1 hour, and then the solvent wasremoved by evaporation to give a solid. The solid was washed with waterto give methyl (S)-2-amino-4-dibenzofuran-2-yl-4-oxo-butyrate as amono-hydrate.

EXAMPLES 18-55

Several invention compounds of Formula I were prepared by combinatorialsynthetic techniques. The general procedure utilized is as follows:

A 0.17 molar stock solution was made by dissolving 2.9 g (8.69 mmol) of2-amino-4-dibenzofuran-2-yl-4-oxo-butyric acid methyl esterhydrochloride (prepared as in Step (a) of Example 6) in 50 mL ofdichloromethane. One milliliter of this solution was added to 38separate vials, along with 70 mg of a morpholino-resin (preparedaccording to Booth R. J. and Hodges J. C., J. Am. Chem. Soc.,1997;119(21):4882-4886). 1.1 Equivalents (0.187 mmol) of the appropriatecarboxylic acid chloride was added to each of the 38 vials. The vialswere sealed and shaken for 54 hours at room temperature. An excess of anamino-resin and an isocyanato-resin (both prepared according to Boothand Hodges, Supra., 1997) was added to each vial, and the vials wereshaken for 16 hours to quench unreacted starting materials. Eachreaction mixture was filtered through a plug of glass wool, and theresins were washed with 2 mL tetrahydrofuran. The filtrate wasevaporated under a stream of nitrogen, and the residue in each vial wasre-dissolved in 1 mL tetrahydrofuran. One milliliter of a 0.315 molaraqueous solution of lithium hydroxide was added to each vial, and theresulting mixtures were again shaken for 16 hours. Each reaction mixturewas washed with diethyl ether, and the aqueous layer was then acidifiedwith 1 molar hydrochloric acid. The products were extracted into ethylacetate, and the solvent was evaporated under a stream of nitrogen for54 hours to leave the desired products. The compounds were analyzed byLC/MS to determine purity and presence of expected ion.

The foregoing reaction is depicted in the following generalized scheme:##STR30##

The following specific invention compounds were prepared by theforegoing combinatorial methodology:

    ______________________________________                                        Example                       MS                                                No. Compound (m + 1)                                                        ______________________________________                                        18        4-Dibenzofuran-2-yl-2-                                                                            478                                                diphenylacetylamino-4-oxo-butyric                                             acid                                                                         19 2-[2-(4-Chloro-phenoxy)- 452                                                acetylamino]-4-dibenzofuran-2-yl-4-                                           oxo-butyric acid                                                             20 4-Dibenzofuran-2-yl-2-(3,4- 456                                             dichloro-benzoylamino-4-oxo-butyric                                           acid                                                                         21 4-Dibenzofuran-2-yl-2-[2-(3,4- 462                                          dimethoxy-phenyl)-acetylamino] -4-                                            oxo-butyric acid                                                             22 4-Dibenzofuran-2-yl-2- --                                                   [(naphthalene-2-carbonyl)-amino] -4-                                          oxo-butyric acid                                                             23 4-Dibenzofuran-2-yl-2-(2,2- 396                                             dimethyl-pentanoylamino)-4-oxo-                                               butyric acid                                                                 24 4-Dibenzofuran-2-yl-4-oxo-2- --                                             [(pyridine-4-carbonyl)-amino]-                                                butyric acid                                                                 25 4-Dibenzofuran-2-yl-4-oxo-2-(3- 414                                         phenyl-acryloylamino)-butyric acid                                           26 2-[(Adamantane-1-carbonyl)-amino]- --                                       4-dibenzofuran-2-yl-4-oxo-butyric                                             acid                                                                         27 4-Dibenzofuran-2-yl-4-oxo-2- --                                             (2,2,3,3,4,4,5,5,6,6,7,7,8,8,8-                                               pentadecafluoro-octanoylamino)-                                               butyric acid                                                                 28 4-Dibenzofuran-2-yl-4-oxo-2-(2- 418                                         phenoxy-acetylamino)-butyric acid                                            29 4-Dibenzofuran-2-yl-2-(4-methyl- 402                                        benzoylamino)-4-oxo-butyric acid                                             30 4-Dibenzofuran-2-yl-4-oxo-2-(2- 432                                         phenoxy-propionylamino)-butyric                                               acid                                                                         31 2-[4-(4-Bromo-phenyl)- 508                                                  butyrylamino]-4-dibenzofuran-2-yl-                                            4-oxo-butyric acid                                                           32 2-(2-Benzyloxy-acetylamino)-4- 432                                          dibenzofuran-2-yl-4-oxo-butyric                                               acid                                                                         33 4-Dibenzofuran-2-yl-2- 396                                                  heptanoylamino-4-oxo-butyric acid                                            34 2-(4-Butyl-benzoylamino)-4- 444                                             dibenzofuran-2-yl-4-oxo-butyric                                               acid                                                                         35 2-(4-Cyano-benzoylamino)-4- 413                                             dibenzofuran-2-yl-4-oxo-butyric                                               acid                                                                         36 2-[(3-Chloro-thiophene-2-carbonyl)- 428                                     amino)-4-dibenzofuran-2-yl-4-oxo-                                             butyric acid                                                                 37 2-(4-Butoxy-benzoylamino)-4- 460                                            dibenzofuran-2-yl-4-oxo-butyric                                               acid                                                                         38 2-(Cyclopropanecarbonyl-amino)-4- 352                                       dibenzofuran-2-yl-4-oxo-butyric                                               acid                                                                         39 2-(4-Chloro-benzoylamino)-4- 422                                            dibenzofuran-2 -yl-4-oxo-butyric                                              acid                                                                         40 4-Dibenzofuran-2-yl-4-oxo-2- --                                             (2,3,4,5,6-pentafluoro-                                                       benzoylamino)-butyric acid                                                   41 4-Dibenzofuran-2-yl-2-[(furan-2- 378                                        carbonyl)-amino]-4-oxo-butyric acid                                          42 2-(4-Bromo-benzoylamino)-4- 466                                             dibenzofuran-2-yl-4-oxo-butyric                                               acid                                                                         43 4-Dibenzofuran-2-yl-2-(4-fluoro- 406                                        benzoylamino)-4-oxo-butyric acid                                             44 4-Dibenzofuran-2-yl-4-oxo-2- 440                                            [(quinoxaline-2-carbonyl)-amino]-                                             butyric acid                                                                 45 4-Dibenzofuran-2-yl-4-oxo-2- 389                                            [(pyridine-2-carbonyl)-amino]-                                                butyric acid                                                                 46 4-Dibenzofuran-2-yl-2-(4-methoxy- 418                                       benzoylamino)-4-oxo-butyric acid                                             47 4-Dibenzofuran-2-yl-2-(3,4- 448                                             dimethoxy-benzoylamino)-4-oxo-                                                butyric acid                                                                 48 4-Dibenzofuran-2-yl-4-oxo-2-(4- 456                                         trifluoromethyl-benzoylamino)-                                                butyric acid                                                                 49 2-(Cyclopentanecarbonyl-amino)-4- 380                                       dibenzofuran-2-yl-4-oxo-butyric                                               acid                                                                         50 2-(Cyclohexanecarbonyl-amino)-4- 394                                        dibenzofuran-2-yl-4-oxo-butyric                                               acid                                                                         51 4-Dibenzofuran-2-yl-4-oxo-2- 394                                            [(thiophene-2-carbonyl)-amino]-                                               butyric acid                                                                 52 4-Dibenzofuran-2-yl-4-oxo-2-(2- --                                          thiophen-2-yl-acetylamino)-butyric                                            acid                                                                         53 2-[(Benzo[1,3)dioxole-5-carbonyl)- 432                                      amino]-4-dibenzofuran-2-yl-4-oxo-                                             butyric acid                                                                 54 4-Dibenzofuran-2-yl-4-oxo-2-(2- 430                                         phenylbutyrylamino)-butyric acid                                             55 4-Dibenzofuran-2-yl-2-[(5-methyl-2- 469                                     phenyl-2H-[1,2,3]triazole-4-                                                  carbonyl)-amino]-4-oxo-butyric acid                                        ______________________________________                                    

The invention compounds have been evaluated in standard in vitro assaysand shown to be potent inhibitors of several matrix metalloproteinaseenzymes. The assays measure the amount by which a test compound reducesthe hydrolysis of a thiopeptolide substrate caused by a matrixmetalloproteinase enzyme. Such assays are described in detail by Ye, etal., in Biochemistry, 1992;31(45):11231-11235).

Thiopeptolide substrates show virtually no decomposition or hydrolysisin the absence of a matrix metalloproteinase enzyme. A typicalthiopeptolide substrate commonly utilized for assays isAc-Pro-Leu-Gly-thioester-Len-Len-Gly-O Et. A 100 μL assay mixture willcontain 50 mM of 2-morpholinoethane sulfonic acid monohydrate (MES, pH6.0) 10 mM CaCl₂, 100 μM thiopeptolide substrate, and 1 mM5,5'-dithio-bis-(2-nitro-benzoic acid (DTNB). The thiopeptolidesubstrate concentration is varied from 10 to 800 μM to obtain Km andKcat values. The change in absorbance at 405 nm is monitored on a ThermoMax microplate reader (molecular Devices, Menlo Park, Calif.) at roomtemperature (22° C.). The calculation of the amount of hydrolysis of thethiopeptolide substrate is based on E₄₁₂ =13600 m⁻¹ cm⁻¹ for theDTNB-derived product 3-carboxy-4-nitrothiophenoxide. Assays are carriedout with and without matrix metalloproteinase inhibitor compounds, andthe amount of hydrolysis is compared for a determination of inhibitoryactivity of the test compounds.

Several representative compounds have been evaluated for their abilityto inhibit various matrix metalloproteinase enzymes. Table I belowpresents inhibitory activity for several invention compounds. In thetable, MMP-1 refers to interstitial collagenase; MMP-2 refers toGelatinase A; MMP-3 refers to stromelysin; MMP-7 refers to matrilysin;and MMP-13 refers to Collagenase 3. Test compounds were evaluated atvarious concentrations in order to determine their respective IC₅₀values, the micromolar concentration of compound required to cause a 50%inhibition of the hydrolytic activity of the respective enzyme.

                  TABLE I                                                         ______________________________________                                        (IC.sub.50) μM                                                               Compound of                                                                   Example No. MMP-1 MMP-2 MMP-3 MMP-7 MMP-13                                  ______________________________________                                         1       100      0.059   0.95   100   2.8                                       2 -- 44.0 78 -- --                                                            3 -- 3.8 33 -- --                                                             4 -- 0.16 1.6 -- --                                                           5 -- 0.32 1.7 -- --                                                           6 -- 0.18 0.91 -- --                                                          7 -- 0.084 0.33 -- --                                                         8 100 0.067 0.29 100 1.3                                                      9 -- 0.51 3.9 -- --                                                          10 -- 1.64 9.4 -- --                                                          11 100 0.36 2.7 100 6.8                                                       12 100 8.8 4.0 -- --                                                          13 -- 0.28 9.2 100 --                                                         14  83 0.017 0.26 100 1.1                                                     15 100 12.5 76.5 100 100                                                      16 100 0.72 6.6 100 2.4                                                       18 100 0.1 0.36 100 1.1                                                       19 100 0.64 3.1 100 46                                                        20 100 1.3 7.0 100 38                                                         21 100 0.32 2.4 100 16                                                        22 100 1.7 6.9 100 28                                                         23 100 0.17 0.76 100 5.6                                                      24 100 3.2 13.0 100 41                                                        25 100 0.11 0.8 100 8.4                                                       26 100 2.1 9.2 100 40                                                         27 100 4.6 18 100 77                                                          28 100 0.18 0.51 100 12                                                       29 100 0.05 0.39 100 5.2                                                      30  45 0.035 0.1 100 2.9                                                      31 100 0.6 5.3 100 25                                                         32 100 0.041 0.33 100 3.5                                                     33 100 0.06 0.64 100 4.7                                                      34 100 0.87 5.4 100 49                                                        35 100 0.11 1.1 100 11                                                        36 100 0.14 1.0 100 13                                                        37 100 0.8 5.0 100 58                                                         38 100 0.09 0.85 100 5.6                                                      39 100 0.15 1.2 100 12                                                        40 100 5.2 28 100 100                                                         41 100 0.14 1.5 100 12                                                        42 100 0.18 1.3 100 17                                                        43 100 0.07 0.66 100 7.1                                                      44 100 0.31 3.5 100 27                                                        45 100 0.11 1.0 100 15                                                        46 100 0.14 1.0 100 11                                                        47 100 0.13 0.87 100 8.8                                                      48 100 0.32 1.5 100 20                                                        49 100 0.08 0.35 100 4.4                                                      50 100 0.15 0.7 100 9.1                                                       51 100 0.16 1.3 100 16                                                        52 100 2.5 7.7 100 48                                                         53 100 0.13 1.0 100 9.2                                                       54 100 0.06 0.25 100 1.5                                                      55 100 1.8 14 100 70                                                        ______________________________________                                    

The compounds were additionally evaluated for their ability to inhibitfull-length collagenase hydrolysis of thiopeptolide substrate (FLC) andfull-length gelatinase B (FLGB) hydrolysis of thiopeptolide. The resultsof representative compounds is given in Table II.

                  TABLE II                                                        ______________________________________                                        (IC.sub.50) μM                                                               Compound of                                                                   Example No. FLC FLGB                                                        ______________________________________                                        1                >100   >100                                                  ______________________________________                                    

The compounds of the present invention can be prepared and administeredin a wide variety of oral and parenteral dosage forms. Thus, thecompounds of the present invention can be administered by injection,that is, intravenously, intramuscularly, intracutaneously,subcutaneously, intraduodenally, or intraperitoneally. Also, thecompounds of the present invention can be administered by inhalation,for example, intranasally. Additionally, the compounds of the presentinvention can be administered transdermally. It will be obvious to thoseskilled in the art that the following dosage forms may comprise as theactive component, either a compound of Formula I or a correspondingpharmaceutically acceptable salt of a compound of Formula I. The activecompound generally is present in a concentration of about 5% to about95% by weight of the formulation.

For preparing pharmaceutical compositions from the compounds of thepresent invention, pharmaceutically acceptable carriers can be eithersolid or liquid. Solid form preparations include powders, tablets,pills, capsules, cachets, suppositories, and dispersible granules. Asolid carrier can be one or more substances which may also act asdiluents, flavoring agents, solubilizers, lubricants, suspending agents,binders, preservatives, tablet disintegrating agents, or anencapsulating material.

In powders, the carrier is a finely divided solid which is in a mixturewith the finely divided active component.

In tablets, the active component is mixed with the carrier having thenecessary binding properties in suitable proportions and compacted inthe shape and size desired.

The powders and tablets preferably contain from five or ten to aboutseventy percent of the active compound. Suitable carriers are magnesiumcarbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin,starch, gelatin, tragacanth, methylcellulose, sodiumcarboxymethylcellulose, a low melting wax, cocoa butter, and the like.The term "preparation" is intended to include the formulation of theactive compound with encapsulating material as a carrier providing acapsule in which the active component, with or without other carriers,is surrounded by a carrier, which is thus in association with it.Similarly, cachets and lozenges are included. Tablets, powders,capsules, pills, cachets, and lozenges can be used as solid dosage formssuitable for oral administration.

For preparing suppositories, a low melting wax, such as a mixture offatty acid glycerides or cocoa butter, is first melted and the activecomponent is dispersed homogeneously therein, as by stirring. The moltenhomogenous mixture is then poured into convenient sized molds, allowedto cool, and thereby to solidify.

Liquid form preparations include solutions, suspensions, and emulsions,for example, water or water propylene glycol solutions. For parenteralinjection, liquid preparations can be formulated in solution in aqueouspolyethylene glycol solution.

Aqueous solutions suitable for oral use can be prepared by dissolvingthe active component in water and adding suitable colorants, flavors,stabilizing, and thickening agents as desired.

Aqueous suspensions suitable for oral use can be made by dispersing thefinely divided active component in water with viscous material, such asnatural or synthetic gums, resins, methylcellulose, sodiumcarboxymethylcellulose, and other well-known suspending agents.

Also included are solid form preparations which are intended to beconverted, shortly before use, to liquid form preparations for oraladministration. Such liquid forms include solutions, suspensions, andemulsions. These preparations may contain, in addition to the activecomponent, colorants, flavors, stabilizers, buffers, artificial andnatural sweeteners, dispersants, thickeners, solubilizing agents, andthe like.

The pharmaceutical preparation is preferably in unit dosage form. Insuch form, the preparation is subdivided into unit doses containingappropriate quantities of the active component. The unit dosage form canbe a packaged preparation, the package containing discrete quantities ofpreparation, such as packeted tablets, capsules, and powders in vials orampoules. Also, the unit dosage form can be a capsule, tablet, cachet,or lozenge itself, or it can be the appropriate number of any of thesein packaged form.

The quantity of active component in a unit dose preparation may bevaried or adjusted from 1 mg to 1000 mg, preferably 10 mg to 100 mgaccording to the particular application and the potency of the activecomponent. The composition can, if desired, also contain othercompatible therapeutic agents.

In therapeutic use as agents for the treatment of multiple sclerosis,atherosclerotic plaque rupture, aortic aneurism, heart failure,restenosis, periodontal disease, corneal ulceration, cancer metastasis,tumor angiogenesis, arthritis, or other autoimmune or inflammatorydisorders dependent upon breakdown of connective tissue, the compoundsutilized in the pharmaceutical method of this invention are administeredat a dose that is effective to inhibit the hydrolytic activity of one ormore matrix metalloproteinase enzymes. The initial dosage of about 1 mgto about 100 mg per kilogram daily will be effective. A daily dose rangeof about 25 mg to about 75 mg per kilogram is preferred. The dosages,however, may be varied depending upon the requirements of the patient,the severity of the condition being treated, and the compound beingemployed. Determination of the proper dosage for a particular situationis within the skill of the art. Generally, treatment is initiated withsmaller dosages which are less than the optimum dose of the compound.Thereafter, the dosage is increased by small increments until theoptimum effect under the circumstance is reached. For convenience, thetotal daily dosage may be divided and administered in portions duringthe day if desired. Typical dosages will be from about 0.1 to about 500mg/kg, and ideally about 25 to about 250 mg/kg.

The following examples illustrate typical formulations provided by theinvention.

EXAMPLE 56

    ______________________________________                                        Tablet Formulation                                                                 Ingredient         Amount (mg)                                           ______________________________________                                        2-N-methylbutyrylamino-4-(6-                                                                      25                                                          chlorodibenzofuran-2-yl)-4-thioxo-                                            butyric acid                                                                  Lactose 50                                                                    Corn starch (for mix) 10                                                      Corn starch (paste) 10                                                        Magnesium stearate (1%)  5                                                    Total 100                                                                   ______________________________________                                    

The dibenzofuranyl butanoic acid, lactose, and corn starch (for mix) areblended to uniformity. The corn starch (for paste) is suspended in 200mL of water and heated with stirring to form a paste. The paste is usedto granulate the mixed powders. The wet granules are passed through aNo. 8 hand screen and dried at 80° C. The dry granules are lubricatedwith the 1% magnesium stearate and pressed into a tablet. Such tabletscan be administered to a human from one to four times a day fortreatment of atherosclerosis, arthritis, and heart failure, includingventricular dilatation.

EXAMPLE 57

    ______________________________________                                        Preparation for Oral Solution                                                     Ingredient            Amount                                              ______________________________________                                        2-Amino-4-carbazol-2-yl-4-hydroxyimino                                                              400       mg                                              butyric acid sodium salt                                                      Sorbitol solution (70% N.F.) 40 mL                                            Sodium benzoate 20 mg                                                         Saccharin 5 mg                                                                Red dye 10 mg                                                                 Cherry flavor 20 mg                                                           Distilled water q.s. 100 mL                                                 ______________________________________                                    

The sorbitol solution is added to 40 mL of distilled water, and thecarbazolyl butyric acid is dissolved therein. The saccharin, sodiumbenzoate, flavor, and dye are added and dissolved. The volume isadjusted to 100 mL with distilled water. Each milliliter of syrupcontains 4 mg of invention compound.

EXAMPLE 58

Parenteral Solution

In a solution of 700 mL of propylene glycol and 200 mL of water forinjection is suspended 20 g of(S)-2-N,N-diethylamino-4-fluoren-3-yl-4-thioxo-butyric acid. Aftersuspension is complete, the pH is adjusted to 6.5 with 1 N sodiumhydroxide, and the volume is made up to 1000 mL with water forinjection. The formulation is sterilized, filled into 5.0 mL ampouleseach containing 2.0 mL, and sealed under nitrogen.

As matrix metalloproteinase inhibitors, the compounds of Formula I areuseful as agents for the treatment of multiple sclerosis. They are alsouseful as agents for the treatment of atherosclerotic plaque rupture,restenosis, periodontal disease, corneal ulceration, treatment of burns,decubital ulcers, wound repair, cancer metastasis, tumor angiogenesis,arthritis, and other inflammatory disorders dependent upon tissueinvasion by leukocytes.

We claim:
 1. A compound of the Formula I ##STR31## wherein: X is O,NOR₉, S, OH, SH, or ##STR32## R₇ and R_(7a), independently arehydrogen,C₁ -C₂₀ alkyl or substituted C₁ -C₂₀ alkyl, (CH₂)₀₋₆ -aryl,dibenzofuran, furan or (CH₂)₀₋₆ -cycloalkyl; R₁ and R₂ independentlyarehydrogen, C₁ -C₂₀ alkyl or substituted C₁ -C₂₀ alkyl, halo, NO₂, CN,CHO, COR₆, COOR₆, SO₃ R₆, OR₆, CONR₄ R₅, (CH₂)₀₋₆ -aryl, dibenzofuran,furan or (CH₂)₀₋₆ -cycloalkyl; R is hydrogen,C₁ -C₂₀ alkyl orsubstituted C₁ -C₂₀ alkyl; aryl is phenyl or substituted phenyl; R₃ ishydroxy,O--C₁ -C₂₀ alkyl or substituted O--C₁ -C₂₀ alkyl, O--(CH₂)₁₋₃aryl, or NHOR₆ ; R₄ and R₅ independently are hydrogen,C₁ -C₂₀ alkyl orsubstituted C₁ -C₂₀ alkyl, C₂ -C₂₀ alkenyl or substituted C₂ -C₂₀alkenyl, (CH₂)₀₋₆ -aryl, (CH₂)₀₋₆ -(O or S)-aryl, dibenzofuran, furan;or one of R₄ and R₅ is hydrogen and the other is:COR₈, CSR₈, CONR₈ R₉,CSNR₈ R₉, COOR₈, COSR₈, ##STR33## S(O)₁ or 2 -C₁ -C₂₀ alkyl orsubstituted alkyl, S(O)₁ or 2 -aryl, dibenzofuran, furan or S(O)₁ or 2-cycloalkyl; Y is--O--, R₈ and R₉ independently arehydrogen, C₁ -C₂₀alkyl or substituted C₁ -C₂₀ alkyl, C₂ -C₂₀ alkenyl or substituted C₂-C₂₀ alkenyl, (CH₂)₀₋₆ -(O or S)₀₋₁ -aryl, dibenzofuran, furan or(CH₂)₀₋₆ -(O or S)₀₋₁ -cycloalkyl; W, W¹, Z, and Z¹ independently areCR₁ ; or a pharmaceutically acceptable salt, isomer, stereoisomer, orand solvate thereof.
 2. A compound of claim 1 having the formula##STR34##
 3. A compound of claim 2 having the formula
 4. A compound ofclaim 3 wherein X is O.
 5. A compound of claim 4 wherein R₃ is C₁ -C₂₀alkoxy.
 6. A compound of claim 4 wherein R₃ is OH.
 7. A compound ofclaim 6 wherein R₄ is H.
 8. A compound of claim 7 wherein R₅ is H. 9.The compound of claim 8 which is(S)-2-amino-4-dibenzofuran-2-yl-4-oxo-butyric acid.
 10. A compound ofclaim 7 wherein R₅ is --COCH₃.
 11. The compound of claim 10 which isS-2-acetylamino-4-dibenzofuran-2-yl-4-oxo-butyric acid.
 12. A compoundof claim 7 wherein R₅ is --COCF₃.
 13. The compound of claim 12 which is(S)-2-(2,2,2-trifluoroacetylamino)-4-dibenzofuran-2-yl-4-oxo- butyricacid.
 14. The compound of claim 12 which is(R)-2-(2,2,2-trifluoroacetylamino)-4-dibenzofuran-2-yl-4-oxo- butyricacid.
 15. A compound of claim 7 wherein R₅ is and R₈ is(CH₂)₀₋₆ -(O orS)₀₋₁ -aryl or dibenzofuran or furan.
 16. A compound of claim 15selected from the group consistingof(S)-2-Benzoylamino-4-dibenzofuran-2-yl-4-oxo-butyric acid;(S)-4-Dibenzofuran-2-yl-4-oxo-2-phenylacetylamino-butyric acid;(S)-4-Dibenzofuran-2-yl-4-oxo-2-(3-phenyl-propionylamino)-butyric acid;(S)-4-Dibenzofuran-2-yl-4-oxo-2-(7-phenyl-heptanoylamino)-butyric acid;(S)-2-[(Biphenyl-4-carbonyl)-amino]-4-dibenzofuran-2-yl-4-oxo-butyricacid;2-[2-(4-Chloro-phenoxy)-acetylamino]-4-dibenzofuran-2-yl-4-oxo-butyricacid; 4-Dibenzofuran-2-yl-2-(3,4-dichloro-benzoylamino4-oxo-butyricacid;4-Dibenzofuran-2-yl-2-[2-(3,4-dimethoxy-phenyl)-acetylarnino]-4-oxo-butyricacid;4-Dibenzofuran-2-yl-2-[(naphthalene-2-carbonyl)-amino]-4-oxo-butyricacid; 4-Dibenzofuran-2-yl-4-oxo-2-(2-phenoxy-acetylamino)-butyric acid;4-Dibenzofuran-2-yl-2-(4-methyl-benzoylamino)-4-oxo-butyric acid;2-[4-(4-Bromo-phenyl)-butyrylamino]-4-dibenzofuran-2-yl-4-oxo-butyricacid; 2-(2-Benzyloxy-acetylamino)-4-dibenzofuran-2-yl-4oxo-butyric acid;2-(4-Butyl-benzoylamino)-4-dibenzofuran-2-yl-4-oxo-butyric acid;2-(4-Cyano-benzoylamino)-4-dibenzofuran-2-yl-4-oxo-butyric acid;2-(4-Butoxy-benzoylamino)-4-dibenzofuran-2-yl-4-oxo-butyric acid;2-(4-Chloro-benzoylamino)4-dibenzofuran-2-yl-4-oxo-butyric acid;4-Dibenzofuran-2-yl-4-oxo-2-(2,3,4,5,6-pentafluoro-benzoylamino)-butyricacid; 4-Dibenzofuran-2-yl-2-[(furan-2-carbonyl)-amino]-4-oxo-butyricacid; 2-(4-Bromo-benzoylamino)-4-dibenzofuran-2-yl-4-oxo-butyric acid;4-Dibenzofuran-2-yl-2-(4-fluoro-benzoylamino)4-oxo-butyric acid;4-Dibenzofuran-2-yl-2-(4-methoxy-benzoylamino)4-oxo-butyric acid;4-Dibenzofuran-2-yl-2-(3,4-dimethoxy-benzoylamino)4-oxo-butyric acid;and 4-Dibenzofuran-2-yl-4-oxo-2-(4-trifluoromethyl-benzoylamino)-butyricacid.
 17. A compound of claim 7 wherein R₅ is ##STR35## and R₈ is C₁-C₁₀ alkyl or substituted C₁ -C₁₀ alkyl.
 18. A compound of claim 17selected from the group consistingof(S)-4-Dibenzofuran-2-yl-4-oxo-2-(octanoyl-amino)-butyric acid;(S)-4-Dibenzofuran-2-yl-4-oxo-2-(dodecanoyl-amino)-butyric acid;4-Dibenzofuran-2-yl-2-diphenylacetylamino-4-oxo-butyric acid;4-Dibenzofuran-2-yl-2-(2,2-dimethylpentanoylamino)-4-oxo-butyric acid;4-Dibenzofuran-2-yl-4-oxo-2-(2,2,3,3,4,4,5,5,6,6,7,7,8,8,8-pentadecafluoro-octanoylamino)-butyricacid; 4-Dibenzofuran-2-yl-4-oxo-2-(2-phenoxypropionylamino)-butyricacid; 4-Dibenzofuran-2-yl-2-heptanoylamino-4-oxo-butyric acid; and4-Dibenzofuran-2-yl-4-oxo-2-(2-phenylbutyrylamino)-butyric acid.
 19. Acompound of claim 7 wherein R₅ is ##STR36## and R₈ is --(CH₂)₀₋₆ -aryl.20. The compound of claim 19 which is(S)-4-Dibenzofuran-2-yl-2-[3-(2,6-diisopropyl-phenyl)-ureido]-4-oxo-butyricacid.
 21. A compound of claim 7 wherein R₅ is S(O)₁ or 2 -C₁ -C₂₀ alkyl.22. The compound of claim 21 which is(S)4-dibenzofuran-2-yl-4-oxo-2-methanesulfonylamino-butyric acid.
 23. Acompound of claim 7 wherein R₅ is S(O)₁ or 2 -aryl.
 24. The compound ofclaim 23 which is(S)-4-dibenzofuran-2-yl-4-oxo-2-(4-methylphenylsulfonylamino)-butyricacid.
 25. A compound of claim 7 wherein R₅ is ##STR37## and R₈ is(CH₂)₀₋₆ -(O or S)₀₋₁ -cycloalkyl.
 26. A compound of claim 25 selectedfrom the group consistingof2-[(Adamantane-1-carbonyl)-amino]-4-dibenzofuan-2-yl-4-oxo-butyricacid; 2-(Cyclopropanecarbonyl-amino)-4-dibenzofuran-2-yl-4-oxo-butyricacid; 2-(Cyclopentanecarbonyl-amino)-4-dibenzofuran-2-yl-4-oxo-butyricacid; and2-(Cyclohexanecarbonyl-amino)-4-dibenzofuran-2-yl-4-oxo-butyric acid.27. A compound of claim 7 wherein R₅ is ##STR38## and R₈ is C₂ -C₂₀alkenyl or substituted C₂ -C₂₀ alkenyl.
 28. The compound of claim 27which is 4-dibenzofuran-2-yl-4-oxo-2-(3-phenyl-acryloylamino)-butyricacid.
 29. A compound of claim 5 which is(S)-2-Benzoylamino-4-dibenzofuran-2-yl-4-oxo-butyric acid methyl ester.30. A compound of claim 3 wherein X is NOR₉.
 31. A compound of claim 30which is(S)-4-dibenzofuran-2-yl-4-hydroxyimino-2-(2,2,2-trifluoroacetylamino)-butyricacid.
 32. A compound of claim 1 having the formula ##STR39##
 33. Apharmaceutical formulation comprising a compound of claim 1 admixed witha diluent, carrier, or excipient therefor.
 34. A method for inhibiting amatrix metalloproteinase enzyme by administering a matrixmetalloproteinase inhibiting amount of a compound of claim
 1. 35. Amethod of treating arthritis comprising administering a matrixmetalloproteinase inhibiting amount of a compound of claim
 1. 36. Amethod of treating restenosis comprising administering a matrixmetalloproteinase inhibiting amount of a compound of claim
 1. 37. Amethod of treating multiple sclerosis comprising administering a matrixmetalloproteinase inhibiting amount of a compound of claim
 1. 38. Amethod of treating atherosclerotic plaque rupture comprisingadministering a matrix metalloproteinase inhibiting amount of a compoundof claim
 1. 39. A method of treating aortic aneurysms comprisingadministering a matrix metalloproteinase inhibiting amount of a compoundof claim
 1. 40. A method of treating heart failure comprisingadministering a matrix metalloproteinase inhibiting amount of a compoundof claim
 1. 41. A method of treating periodontal disease comprisingadministering a matrix metalloproteinase inhibiting amount of a compoundof claim
 1. 42. A method of promoting wound healing comprisingadministering a matrix metalloproteinase inhibiting amount of a compoundof claim 1.